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A Multiplex PCR Assay for Differential Identification of Wild-type and Vaccine Strains of Mycoplasma gallisepticum

Mycoplasma gallisepticum (MG) can cause respiratory disease in chickens and result in serious economic losses in the chicken industry. The use of live vaccines has been a favorable option for the control of MG infection in multi-age commercial layers and broiler breeders. There are three live vaccin...

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Autores principales: Kang, Sung-Il, Lee, O-Mi, Lee, Hye-Jin, Kwon, Yong-Kuk, Chae, Myeong Ju, Jeong, Ji-Yeon, Kang, Min-Su
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9865210/
https://www.ncbi.nlm.nih.gov/pubmed/36678459
http://dx.doi.org/10.3390/pathogens12010111
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author Kang, Sung-Il
Lee, O-Mi
Lee, Hye-Jin
Kwon, Yong-Kuk
Chae, Myeong Ju
Jeong, Ji-Yeon
Kang, Min-Su
author_facet Kang, Sung-Il
Lee, O-Mi
Lee, Hye-Jin
Kwon, Yong-Kuk
Chae, Myeong Ju
Jeong, Ji-Yeon
Kang, Min-Su
author_sort Kang, Sung-Il
collection PubMed
description Mycoplasma gallisepticum (MG) can cause respiratory disease in chickens and result in serious economic losses in the chicken industry. The use of live vaccines has been a favorable option for the control of MG infection in multi-age commercial layers and broiler breeders. There are three live vaccines, including ts-11, 6/85, and F strain, that have been commonly used in various parts of the world, including South Korea. The definitive diagnosis of the infection, therefore, requires the differentiation of wild-type field strains of MG from the vaccine strains used. Thus, we aimed to develop a novel multiplex PCR assay to discriminate between vaccine strains (ts-11, 6/85, and F strain) and wild-type field strains of MG isolated from infected chickens. We designed four novel primer sets that are each specific to MG species, ts-11, 6/85, and F strain. The multiplex PCR assay using the primer sets differentially identified wild-type and vaccine strains of MG but did not detect other avian bacteria. The detection limit of this assay was 250 fg/μL of genomic DNA of each strain tested. In addition, this assay was applied to 36 MG strains isolated from chickens over the past 20 years in South Korea. As a result, the assay identified 22 wild-type strains and 14 vaccine strains. Consequently, the novel multiplex PCR assay can discriminate between vaccine and wild-type field strains of MG and could be a valuable tool for the diagnosis of MG infection in MG-vaccinated chicken flocks.
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spelling pubmed-98652102023-01-22 A Multiplex PCR Assay for Differential Identification of Wild-type and Vaccine Strains of Mycoplasma gallisepticum Kang, Sung-Il Lee, O-Mi Lee, Hye-Jin Kwon, Yong-Kuk Chae, Myeong Ju Jeong, Ji-Yeon Kang, Min-Su Pathogens Article Mycoplasma gallisepticum (MG) can cause respiratory disease in chickens and result in serious economic losses in the chicken industry. The use of live vaccines has been a favorable option for the control of MG infection in multi-age commercial layers and broiler breeders. There are three live vaccines, including ts-11, 6/85, and F strain, that have been commonly used in various parts of the world, including South Korea. The definitive diagnosis of the infection, therefore, requires the differentiation of wild-type field strains of MG from the vaccine strains used. Thus, we aimed to develop a novel multiplex PCR assay to discriminate between vaccine strains (ts-11, 6/85, and F strain) and wild-type field strains of MG isolated from infected chickens. We designed four novel primer sets that are each specific to MG species, ts-11, 6/85, and F strain. The multiplex PCR assay using the primer sets differentially identified wild-type and vaccine strains of MG but did not detect other avian bacteria. The detection limit of this assay was 250 fg/μL of genomic DNA of each strain tested. In addition, this assay was applied to 36 MG strains isolated from chickens over the past 20 years in South Korea. As a result, the assay identified 22 wild-type strains and 14 vaccine strains. Consequently, the novel multiplex PCR assay can discriminate between vaccine and wild-type field strains of MG and could be a valuable tool for the diagnosis of MG infection in MG-vaccinated chicken flocks. MDPI 2023-01-09 /pmc/articles/PMC9865210/ /pubmed/36678459 http://dx.doi.org/10.3390/pathogens12010111 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kang, Sung-Il
Lee, O-Mi
Lee, Hye-Jin
Kwon, Yong-Kuk
Chae, Myeong Ju
Jeong, Ji-Yeon
Kang, Min-Su
A Multiplex PCR Assay for Differential Identification of Wild-type and Vaccine Strains of Mycoplasma gallisepticum
title A Multiplex PCR Assay for Differential Identification of Wild-type and Vaccine Strains of Mycoplasma gallisepticum
title_full A Multiplex PCR Assay for Differential Identification of Wild-type and Vaccine Strains of Mycoplasma gallisepticum
title_fullStr A Multiplex PCR Assay for Differential Identification of Wild-type and Vaccine Strains of Mycoplasma gallisepticum
title_full_unstemmed A Multiplex PCR Assay for Differential Identification of Wild-type and Vaccine Strains of Mycoplasma gallisepticum
title_short A Multiplex PCR Assay for Differential Identification of Wild-type and Vaccine Strains of Mycoplasma gallisepticum
title_sort multiplex pcr assay for differential identification of wild-type and vaccine strains of mycoplasma gallisepticum
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9865210/
https://www.ncbi.nlm.nih.gov/pubmed/36678459
http://dx.doi.org/10.3390/pathogens12010111
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